tbuckley/cabot-search huggingface dataset:
Abstract P2164: Expression Of Engineered Bacterial Sodium Channel Improves Cardiomyocyte Contractility
Authors: Tianyu Wu, Binjie Li, James Koconis, Abhishek Bhattacharjee, Lei Ye, Nenad Bursac
Journal: Circulation Research (2023)

Introduction: Our recent studies in non-human primates have demonstrated adeno-associated virus (AAV) delivery of prokaryotic sodium channel (BacNa v ) can improve the contractile function of infarcted hearts. We hypothesized that BacNa v expression increases cardiomyocyte (CM) contractile function by indirectly augmenting Ca 2+ transient amplitude via increase in sarcoplasmic reticulum (SR) Ca 2+ stores. Methods: We developed an in vitro model of ischemia-reperfusion (I/R) injury in engineered heart tissues (cardiopatches) made from human iPSC-derived CMs (hiPSC-CMs). Prior to formation, tissues were transduced with lentivirus conferring a doxycycline (Dox)-inducible expression of BacNa v . Upon I/R injury, the tissues were treated with either vehicle or Dox for 48h followed by force testing and Ca 2+ imaging. The effect of BacNa v expression on Ca 2+ handling was additionally studied using neonatal rat ventricular myocytes (NRVMs). Results: The tissue injury from I/R was evident from increases in cleaved-caspase 3 area (0.25% vs 1.53%), ROS generation (1.2 fold), LDH release (4.1 fold), and percent dead cells (0.97% vs 4.67%). Immediately following the I/R injury, tissues showed decrease in contractile force (2.78mN vs 0.74mN) and conduction velocity (20.7cm/s vs 7.9cm/s). At 72hr post-injury, BacNa v -expressing tissues exhibited significantly higher contractile force (1.67mN vs 1.32mN) and Ca 2+ transient (1.8 fold) amplitudes compared to the vehicle control. Furthermore, in uninjured NRVM monolayers, BacNa v expression resulted in higher Ca 2+ transient amplitude (1.31 fold), as well as increased SR calcium stores (1.3 fold) and slower sodium/calcium exchanger (NCX) extrusion rate (1.03s -1 vs 0.82s -1 ) in the presence of caffeine. Blocking NCX with 10M ORM-10962 eliminated the difference in Ca 2+ transient amplitude between BacNa v -expressing and control CMs. Conclusion: Our results suggest BacNa v expression augments CM contractility and Ca 2+ transient amplitude, at least in part through the suppression of Ca 2+ efflux mode of the NCX. In addition to its antiarrhythmic effects, BacNa v gene delivery may represent an effective therapeutic strategy to improve cardiac contractility in congestive heart failure.
Abstract P1074: <i>In Vitro</i> Stimulation-conduction Platforms To Mature Induced Pluripotent Stem Cell-derived Cardiomyocytes
Authors: Suh Hee Cook, Jack Twiddy, Michael A Daniele, Jessica M Gluck
Journal: Circulation Research (2023)

Figure 1. Scaffold (i) and bioreactor (ii) viability for iPSC-CM conduction-stimulation platform.Heart disease is the leading cause of death worldwide. Cardiac tissue engineering offers various solutions, frequently employing induced pluripotent stem cell-derived cardiomyocytes (iPSC-CMs). iPSC-CMs can be expanded limitlessly and used autologously; however, they are immature compared to healthy, adult cardiomyocytes in vivo . We are interested in conductive scaffolds to mature iPSC-CMs by facilitating gap-junctional coupling in the presence of electrical stimulation. We have previously endowed electrospun scaffolds with high conductivity by incorporating carbon nanotubes (CNTs). We differentiated iPSCs into iPSC-CMs with a Wnt inhibitor-directed protocol. We confirmed CNT scaffold biocompatibility by seeding iPSC-CMs onto scaffolds with and without CNTs; 24 hours later, a live/dead assay indicated scaffolds exhibited no cytotoxicity (Fig. 1i). We also confirmed that the application of voltage-controlled electrical stimulation (square wave, 5 ms period, 20% duty cycle) for 5 minutes at magnitudes of 0, 0.1, 1, and 10 V did not introduce cytotoxicity (Fig. 1ii). Furthermore, application of 0.1 V resulted in earlier ultrastructure development and beating. We have developed an upgraded bioreactor which we will use to probe the role of electrical signals in embryonic cardiac development and to explore the ability of CNT scaffolds to electrophysiologically mature iPSC-CMs in the presence of electrical pacing. If successful, this could pave the way for advances in cellular pacemaking, physiologically relevant models, and cardiac tissue regeneration.
Abstract P2162: Controlled Porous Structures Of Nitric Oxide-releasing Inhaler For Pulmonary Arterial Hypertension Vascular Reverse Remodeling
Authors: Yoogyeong Oh, Jinkee Hong, Se Yong Jung
Journal: Circulation Research (2023)

Pulmonary arterial hypertension (PAH) is an incurable disease characterized by elevated blood pressure in the pulmonary artery. Nitric oxide (NO) is a potent regulator of hemodynamics and macrophage reprogramming. However, the therapeutic effects of NO depend on its concentration and its half-life is very short. Pulmonary delivery has advantages such as the ability to escape first-pass metabolism and the large absorptive surface area. In this study, we prepared NO-releasing inhalers with controlled porous structures to modulate the deposition in the respiratory system. NO-releasing inhalers with micro-sized pores showed improved nebulization efficiency than nano-sized ones. Furthermore, the cGMP expression and macrophage polarization were evaluated to confirm the vasodilatory and anti-inflammatory effects of NO. The customized nebulizer system was prepared for efficiency test of NO-MPs. The fluorescence intensity of Nile red was analyzed using photoluminescence spectroscopy to confirm the amounts of NO-MPs on the slide glass. As with confocal images, 0.8k BPEI/NO-MPs could arrive the lower region than 25k BPEI/NO-MPs. NO produced by endothelial nitric oxide synthase (eNOS) induce vasodilation by increasing cyclic guanosine monophosphate (cGMP) in vascular smooth muscle cells. Hence, the delivery of NO mimicking the physical production of the endothelium in range of 0.05-0.4 nmol cm -2 min -1 is required to restore vascular homeostasis as NO has a concentration-dependent effect on cells. Therefore, BPEI0.8k/NO-MPs release 0.277 nmol cm -2 min -1 of NO that mimics the environment in healthy endothelium, while BPEI25k/NO-MPs showed a higher concentration of NO release, 0.548 nmol cm -2 min -1 . BPEI0.8k/NO-MPs increased the synthesis of cGMP in cells, whereas BPEI25k/NO-MPs presented adverse effects due to an initial burst release. When cell populations were quantified, BPEI0.8k/NO-MPs increased the number of M2-like cells while decreasing the number of M1-like cells. Furthermore, when we analyzed proteins and gene levels of cells, anti-inflammatory factors increased in BPEI0.8k/NO-MPs treatment groups. In conclusion, NO-releasing inhalers could deliver NO to deep lungs and has the potential for pulmonary vascular recovery.
Abstract P3125: Big Data Evaluation Of Cardiac Remodeling Trajectories After Myocardial Infarction In Male And Female Mice
Authors: Jocelyn R Rodriguez-Paar, Jop van Berlo, Merry L Lindsey, Upendra Chalise
Journal: Circulation Research (2023)

Female mice display higher survival rates by day (D) 7 after myocardial infarction (MI) compared to their male counterparts. We used big data analysis of the mouse Heart Attack Research Tool (mHART) 2.0 database to identify sex similarities and differences in cardiac wound healing after MI. The database includes echocardiogram and necropsy analysis from n=1,746 (1,146M and 600F) C57BL/6J wild-type adult mice (age 2-9 months). The extent of cardiac dilation over the first week after MI (calculated as fold-change of average D0 value) was parallel in males and females on all days except MI D5. At D5, males displayed 22% higher end systolic volume and 17% higher end diastolic volume than females (both p&lt;0.0001). D5 infarct size did not explain the dilation or survival differences (53±1% in males and 50±2% in females, p&gt;0.99). Analysis of extracellular matrix (ECM) gene expression data for 84 ECM genes (by Qiagen array) from no MI D0 left ventricle (n=40, 24M/16F) and MI D5 (n=28, 21M/7F) infarct tissue revealed molecular signaling differences by sex. Compared to D0, MI D5 showed 70 ECM genes differentially expressed (all p&lt;0.05). MI D5 showed 38 genes differentially expressed by sex (all p&lt;0.05 by t-test), of which 20 were higher and 18 were lower in males. The top 3 genes ranked by variable important scores were Timp3, Col4a2, and Mmp7 , and all were increased in males. The most enriched pathway represented was focal adhesion in males and matrix metalloproteinases in females. Focal adhesion was enriched in both males and females at MI D5, albeit through upregulation of different gene profiles; Itgb1, Col3a1, Col4a1/2 in males and Lamb3, Lama1, and Itgax in females. Overall, our results indicate that male and female mice show unique cardiac remodeling trajectories with notable divergence at D5, leading to higher early survival.
Abstract P1066: Engineering Of Ferritin Overexpressing Human Induced Pluripotent Stem Cells For In Vivo Tracking Of Grafts Using Magnetic Resonance Imaging
Authors: Yura Son, Wuqiang Zhu
Journal: Circulation Research (2023)

Introduction: Transplantation of human induced pluripotent stem cells-derived cardiomyocytes (hiPSC-CMs) emerges as a promising approach for the treatment of heart diseases. However, it is an unmet challenge to precisely localize the hiPSC-CMs upon transplantation and quantify the graft size in the live large animals. To address this issue, we established a new approach to track grafted CMs in large animals using magnetic resonance imaging detectable reporter molecules, which allows the assessment of graft size in live animals at different time points. Methods: We employed a CRISPR/Cas9 system as a virus-free approach to overexpress ferritin in hiPSCs. Results: hiPSC cells were transfected with ferritin heavy chain CRISPR activation plasmid for 24 hours. Before antibiotic selection, we established a kill curve to determine the appropriate antibiotic concentration. Based on the kill curve, we conducted antibiotic selection using puromycin for 7 days and isolated transfected colonies for further culture. Next, we treated various concentrations of iron chloride (0-100 μM) to the FHC transfected hiPSC for 4 hours to examine a proper concentration of iron loading for MRI detection. There was no significant difference in cell viability between 0 and 25 μM of iron chloride. In addition, we confirmed transfection efficiency with higher ferritin expression in transfected cells compared to controls (non-transfected wild-type hiPSC). Following, we plan to differentiate ferritin overexpressing hiPSC into cardiomyocytes, inject the cells into porcine hearts, and conduct magnetic resonance imaging to detect the engrafted transgenic cells in live animals. Conclusion: The ferritin-expressing hiPSC line has the potential to provide a reliable measurement of the graft size in large animals.
Abstract P3123: Sex-related Differences In Cardiac Biomarkers And Ventricular Fibrosis In A Minipig Model Of Hypertrophic Cardiomyopathy
Authors: Aileen Castro Coronado, Gail E Geist, Trisha Smit, Christopher Rogers, Frank Rohret, Carlos L Del Rio, Ricardo Garcia, David M Ryba
Journal: Circulation Research (2023)

Hypertrophic Cardiomyopathy (HCM) is characterized by ventricular hypertrophy and stiffness resulting in part from interstitial fibrosis. While associated with autosomal dominant mutations in sarcomere proteins, it has been reported that males and females may have different clinical presentations and disease courses. We have previously shown that minipigs carrying the R403Q mutation in MYH7 demonstrate a phenotype that recapitulates human non-obstructive HCM with increased hypertrophy, markers of cardiac stress, and diastolic dysfunction. Under the hypothesis that sex-dependent differences in cardiac physiology can independently modulate disease phenotype, we evaluated ventricular fibrosis and cardiac biomarkers in both male and female mutant pigs. Purpose-bred Yucatan R403Q MYH7 heterozygous mutant minipigs of either sex (M: n=8, F: n=6) and wild-type littermates (M: n=4, F: n=14) were used; for the purposes of these comparisons, cloned male pigs were also included (R403Q: 8). Males were castrated, and all pigs received soybean-based feed. Circulating levels of both cardiac troponin T (cTnT) and brain natriuretic peptide (BNP), markers of cardiac injury and stress, were assessed. Interstitial fibrosis was assessed in left ventricular free wall (LV-FW) and apical tissue samples utilizing picrosirius red staining and semi-automated quantification (HALO). Male HCM pigs died earlier (&lt;9M of age, P&lt;0.05) and had higher serum biomarkers than females (cTnT levels (M: 143±17 vs F: 44±7 ng/L, P&lt;0.05; BNP 1536±335 vs 4881±414 ng/L, P&lt;0.05) Consistent with the higher cTnT levels, male HCM pigs had greater fibrosis areas than females in both LV-FW (11±3% vs 7±2%, P&lt;0.05) and apical regions (13±1% vs. 8±2%, respectively, P&lt;0.05). All HCM pigs had significantly increased fibrotic area vs WT controls (R403Q: 10±3% vs WT: 3±2%, P&lt;0.05). Our findings demonstrate that despite a common genotype, female and male carriers of the HCM pathogenic R403Q mutation present different disease phenotypes, with castrated males showing greater fibrotic burden. Future studies to understand the mechanistic drivers of these differences should not only help elucidate the role of sex in HCM disease progression, but also the development of sex-informed therapies.
Abstract P2048: New Insights Into The Role Of The Glycocalyx Components In Heart Failure Induced By Pressure Overload.
Authors: Lakshmi Narasimha Thota, Simone Potje, Lusiane Bendhack, Joao Carvalho-de-Souza, Andreia Chignalia
Journal: Circulation Research (2023)

Heart failure (HF) is a complex disease with multifactorial etiology. The degradation of the glycocalyx is a process thought to contribute to the progression of HF. The relative individual contribution of glycocalyx components to HF progression is unclear. Previous studies have shown that the expression of glypican 1 (Gpc1), a glycocalyx component, is reduced after aortic banding. Herein, we sought to determine if Gpc1 participates in the progression of HF induced by pressure overload and the associated mechanisms. Methods: Male, 5-8 weeks old, WT and Gpc1 knockout mice (Gpc1KO) were submitted to transaortic constriction (TAC) or sham procedure. Heart function was assessed by echocardiography; heart remodeling was assessed by histological analysis; heparan sulfate and glypican 1 content was assessed by immunofluorescence; the expression of ANP and myosin was assessed by immunoblotting. Results: Gpc1KO showed preserved heparan sulfate content in heart sections. Gpc1KO showed cardiac hypertrophy as evidenced by increased left ventricle mass (173.75mg vs. 123.96 mg in WT; p=0.004; n=10), left ventricle wall thickness (1.43mm vs . 1.13mm in WT; p=0.033; n=10), intraventricular septal end-diastolic thickness (1.83mm vs. 1.62mm in WT; p=0.001; n=11) and decreased cardiomyocyte cross-sectional area (203.86μm2 vs. 248.44μm2; p&lt;0.0001; n=5). This was associated with increased stroke volume (52.89±2.1μL vs 46.42±2.0 μL; p=0.038, n=10), cardiac output (26.39 mL/min vs. 20.32 mL/min in WT), calcium deposits, ANP (0.86±0.02 vs. 0.57± 0.03; p&lt;0.0001; n=5) and myosin (0.71±0.1 vs. 0.50±0.01 in WT; p=0.03; n=5) content in the heart. Gpc1KO were prone to TAC-induced HF as shown by decreased ejection fraction (34.37% vs . 58.41% in WT; p=0.047; n=3) and stroke volume (35.24μL vs. 44.60 μL in WT; p=0.05, n=3) 3 weeks after TAC. Moreover, Gpc1KO showed increased mortality rate, not surviving 5 weeks post-TAC. Conclusions: Gpc1KO shows basal cardiac remodeling associated with changes in calcium dynamics and increased contractility. The increased susceptibility of Gpc1KO to TAC-induced HF seems to be independent of glycocalyx degradation. Further studies are required to understand the mechanisms whereby Gpc1 regulates the progression of TAC-induced HF.
Abstract P1072: Microneedle Patch Loaded With Il-4 Macroparticles Facilitates Heart Repair Through Macrophages Mediated Cardiomyocyte Phenotype Transition
Authors: Ke Huang
Journal: Circulation Research (2023)

Ischemic heart disease remains the leading cause of mortality worldwide, but significant progress has been made in optimizing the treatment of acute myocardial infarction (MI) with percutaneous coronary intervention techniques. However, the increased pool of post-MI survivors has led to a higher prevalence of heart failure (HF) secondary to coronary artery disease, making it a major public health issue. Monocytes recruited from circulation differentiate into pro-inflammatory macrophages (M1-like) responsible for post-MI inflammation and cellular debris removal. As a unique Th2 cell cytokine, Interleukin 4 (IL-4) has been widely established as a powerful agent for the polarization of anti-inflammatory macrophages that promote tissue repair. However, there are limitations such as local and targeted delivery have hampered the successful translation of IL-4 therapy. In this study, we created an IL-4 microparticle-loaded microneedle patch (IL-4 MP-MN) for localized delivery of IL-4 to the MI region and tested the therapeutic effects in a rat model of MI. Our preliminary data suggested that IL-4 MP-MN treatment promoted cardiomyocyte proliferation and induced an anti-inflammatory microenvironment, which further led to a cardiomyocyte phenotype shift from "self-rescue" to "assisted-rescue" mode. Additionally, we successfully translated the therapeutic effects of IL-4 MP-MN in a porcine model of MI for testing therapeutic feasibility, safety, and efficacy.
Abstract P3139: Unsupervised Anomaly Detection For Identifying Arrhythmogenic Rhythms In Atrioventricular Block Hearts Using Deep Convolutional Autoencoders
Authors: Hyeong Kyun Park, Young Hoon Son, Nam K Kim, Jeongin Jang, Christina Sheng, Dahim Choi, Junbeom Park, Hee Cheol Cho, Sung-Jin Park
Journal: Circulation Research (2023)

Atrioventricular block (AVB) is a condition that can cause delayed or absent impulse conduction and increase the risk of arrhythmic complications such as atrial fibrillation (AF), premature ventricular contraction (PVC), and ventricular tachycardia (VT). Therefore, long-term monitoring of the heart's AVB rhythms and the ability to identify and predict abnormal rhythms is crucial for clinical decision-making in AVB patients. However, current tools to analyze irregular AVB rhythms and identify anomalies are limited. While supervised deep learning algorithms have been effective for anomaly detection during regular ordered sinus rhythm, they lack practicality for analyzing irregular AVB ECG rhythms. To address this challenge, we developed an unsupervised anomaly detection approach using deep convolutional autoencoder (CAE) models. We created three different CAE models consisting of either residual blocks (ResBlocks) or standard and transpose convolutional layers (TransConv1Ds) to identify anomalies in AVB ECG data. We acquired 30 days of continuous ECG data from seven porcine AVB hearts with complete AV node blockage and optimized the models accordingly. Both ResBlocks- and TransConv1Ds-based CAE models demonstrated high sensitivity and specificity to identify AF, PVC, and VT compared to a PCA-based traditional autoencoder model, with a true positive rate of 92-99% and a true negative rate of 85-92%. The ResBlocks-based CAE models showed high robustness across different ECG data acquired at different days and various porcine models compared to a TransConv1D-based model. As a case application of our anomaly detection approach, we applied the CAE to evaluate the efficacy and toxicity of TBX18-based biological pacemakers for treating AVB hearts. Our CAE analysis revealed that injecting these pacemakers at the ventricular high septum increased the occurrence of AF and PVC compared to the control group, suggesting that the rhythms from the biological pacemakers competed with the ventricular rhythms of the AVB heart. Moreover, the CAE demonstrated high sensitivity, specificity, and robustness in identifying anomalies from human AVB ECG rhythms, indicating the potential for unsupervised algorithms to improve AVB management and treatment.
Abstract P1185: Spatial Transcriptome Profiling In The Mouse Heart With Single-Cell Resolution
Authors: Yuening Liu, Annabelle Coles, Abigail Neininger-Castro, Julia Rune, Richard T Lee, Pallav Kosuri
Journal: Circulation Research (2023)

Background: The spatial organization of the diverse cells in the heart shapes biological function. For instance, during the inflammatory and fibrotic processes that are central to heart disease, many cells act mainly within their local microenvironment. Recent studies using single-cell RNA sequencing (scRNA-seq) have provided a detailed list of the cell types and states involved in development, homeostasis and in cardiovascular diseases. However, scRNA-seq requires tissue dissociation, which removes spatial information and thereby limits our ability to determine how cells interact with each other to maintain health or cause disease. As a consequence, the precise spatial organization of cell types and states in the mammalian heart remains unknown. We sought to address this knowledge gap by using multiplexed error-robust fluorescence in situ hybridization (MERFISH), a method that enables simultaneous identification and localization of single transcripts from hundreds of different genes in intact tissue sections. Methods/Results: We adapted MERFISH and developed a new single-cell image segmentation method to creating a single-cell atlas of the mouse heart. First, we designed and constructed a 128-gene MERFISH probe library including cell type marker genes and also genes covering a broad range of cardiac signaling pathways. We then optimized sample preparation, including cell boundary staining and tissue clearing. Once optimized, we used this approach to image 10 μm-thick sections from healthy mouse heart, resolving tens of thousands of cells and millions of transcripts. Our data thus constitute the first single-cell spatial transcriptome atlas of the mouse heart. Conclusion: Through our newly developed cardiac MERFISH platform, we identified tens of thousands of cells and labeled millions of individual RNA transcripts in intact sections of mouse heart. Our cellular transcriptome profile maps enable new insights into how the organization of the heart shapes its biological function.